CircRERE调节miR-128-3p/WEE1轴促进人多发性骨髓瘤细胞系增殖、迁移和侵袭

doi:10.16352/j.issn.1001-6325.2024.02.0210

摘要/Abstract

摘要： 目的探究环状RNA RERE(circRERE)调节微小RNA(miR)-128-3p/丝氨酸/苏氨酸蛋白激酶(WEE1)轴对人多发性骨髓瘤(MM)细胞系增殖、迁移和侵袭的影响。方法体外培养从健康受试者外周血中分离的正常浆细胞(nPCs)和人MM细胞系(U266、RPMI-8226、NCI-H929、LP-1)。RT-qPCR检测circRERE、miR-128-3p表达;Western blot检测WEE1表达。转染RPMI-8226细胞后,将其分为对照组(control)(未进行转染)、sh-circRERE组、sh-NC组、miR-128-3p inhibitor组、inhibitor-NC组、sh-circRERE+inhibitor-NC组、sh-circRERE+miR-128-3p inhibitor组,噻唑蓝(MTT)实验和5-乙炔基-2’脱氧尿嘧啶核苷(EdU)免疫荧光染色检测细胞增殖;划痕愈合实验、Transwell小室法检测细胞迁移、侵袭;双荧光素酶报告基因实验证实miR-128-3p与circRERE或WEE1的靶向作用关系。结果与nPCs比较,MM细胞(RPMI-8226、U266、NCI-H929、LP-1)中circRERE、WEE1表达增加,miR-128-3p表达减少(P＜0.05);与对照组比较,sh-circRERE组RPMI-8226细胞增殖率、EdU阳性细胞比例、划痕面积愈合率、侵袭数均减少(P＜0.05),miR-128-3p inhibitor组则相反(P＜0.05);miR-128-3p inhibitor降低了sh-circRERE对RPMI-8226细胞增殖、迁移、侵袭的影响(P＜0.05)。双荧光素酶实验显示circRERE/miR-128-3p、miR-128-3p/WEE1存在靶向作用关系。结论 CircRERE能够促进RPMI-8226细胞增殖、迁移和侵袭,可能通过对miR-128-3p发挥海绵效应以增加WEE1表达而实现。

关键词: circRERE, miR-128-3p/WEE1轴, 多发性骨髓瘤细胞, 增殖, 迁移

Abstract: Objective To investigate the impacts of circular RNA RERE (circRERE) on proliferation, migration and invasion of human multiple myeloma (MM) cell lines by regulating microRNA (miR)-128-3p/serine/threonine protein kinase (WEE1) axis. Methods Normal plasma cells (nPCs) isolated from peripheral blood of healthy subjects and human MM cell lines(U266, RPMI-8226, NCI-H929, LP-1) were cultured. The expressions of circRERE and miR-128-3p were detected by RT-qPCR. The expression of WEE1 was detected by Western blot. After transfection, RPMI-8226 cells were divided into control group (without transfection), sh-circRERE group, sh-NC group, miR-128-3p inhibitor group, inhibitor-NC group, sh-circRERE+inhibitor-NC group and sh-circRERE+miR-128-3p inhibitor group. MTT assay and 5-ethynyl-2'-deoxyuridine (EdU) immunofluorescence staining microscopy were used to detect the proliferation. Migration and invasion were examined by scratch healing experiment as well as Transwell chamber assay. The targeting relationship between miR-128-3p and circRERE or WEE1 was verified though dual-luciferase reporter gene assay. Results Compared with nPCs, the expressions of circRERE and WEE1 in MM cells (RPMI-8226, U266, NCI-H929, LP-1) were increased, and the expression of miR-128-3p was decreased (P＜0.05). Compared with the control group, the proliferation rate, proportion of EdU positive cells, the healing rate of the scratch trauma and the number of invasion of the RPMI-8226 cells in sh-circRERE group were decreased(P＜0.05). However,miR-128-3p inhibitor group showed an opposite result and the difference was statistically significant (P＜0.05). miR-128-3p inhibitor significantly inhibited the effects of sh-circRERE on the proliferation, migration and invasion of RPMI-8226 cells (P＜0.05). Dual luciferase reporter gene experiments showed that circRERE/miR-128-3p and miR-128-3p/WEE1 had a targeting relationship. Conclusions CircRERE may promote proliferation, migration and invasion of RPMI-8226 cells, potentially with a mechanism of regulating miR-128-3p/WEE1 axis.

Key words: circRERE, miR-128-3p/WEE1 axis, multiple myeloma cells, proliferation, migration

中图分类号:

R733.3

方媛, 李轶, 王玮. CircRERE调节miR-128-3p/WEE1轴促进人多发性骨髓瘤细胞系增殖、迁移和侵袭[J]. 基础医学与临床, 2024, 44(2): 210-218.

FANG Yuan, LI Yi, WANG Wei. CircRERE promotes proliferation, migration and invasion of human multiple myeloma cell lines by regulating miR-128-3p/WEE1 axis[J]. Basic & Clinical Medicine, 2024, 44(2): 210-218.

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